Integrated platform for selective microfluidic particle processing

Abstract

The instrument for processing microscopic particles may include a combination of modules integrated into a functional system. The system may be composed of synchronized subsystems including pneumatic modules, fluorometers, multi-wavelength lasers, dual view microscopy, high-voltage generators and other subsystems designed to analyze and control microfluidic processes occurring on a chip. The system performs the combined functions of microfluidic particle generation, analysis, and selection, making it possible for the user to create custom integrated workflows for molecular and cell biology processing. Creating the integrated platform requires an innovative and multi-disciplinary approach for module mechanical integration, electronic synchronization and user interface design for controlling diverse microfluidic processes at high speed.

We claim: 
     
       1. An optics module for use with a microfluidic chip comprising particles flowing from a detection zone to a selection zone, the optics module comprising:
 a first image sensor that defines a first image sensor optical path that intersects the selection zone, wherein the first image sensor optical path comprises an objective with a numeric aperture of less than 0.3; and wherein the first image sensor captures images of the particles in the selection zone; 
 a plurality of lasers that define a laser optical path and intersects the detection zone, wherein the plurality of lasers induces fluorescence excitation in the particles; 
 a fluorescence detector assembly that defines a fluorescence detector optical path that intersects the detection zone, wherein a portion of the fluorescence detector optical path is along the laser optical path; and wherein the fluorescence detector assembly detects fluorescence excitation in the particles; 
 a second image sensor that defines a second image sensor optical path that intersects the detection zone, wherein a portion of the second image sensor optical path is along the laser optical path; wherein the second image sensor optical path comprises an objective with a numeric aperture of greater than 0.3; wherein the second image sensor captures images of the particles in the detection zone; and wherein the first image sensor and the second image sensor capture and transmit at least 2000 image frames per second with a latency time of less than 100 microseconds. 
 
     
     
       2. The optics module of  claim 1 , wherein the fluorescence detector assembly comprises a plurality of fluorescence detectors and a plurality of filters that detect the fluorescence excitation in the particles at a plurality of wavelengths. 
     
     
       3. The optics module of  claim 2 , wherein the plurality of wavelengths is selected from a group consisting of: 405 nm, 452 nm, 525 nm, 600 nm, and 680 nm. 
     
     
       4. The optics module of  claim 1 , wherein the plurality of lasers emit laser light at a plurality of wavelengths. 
     
     
       5. The optics module of  claim 4 , wherein the plurality of wavelengths is selected from a group consisting of: 405 nm, 488 nm, 561 nm, and 638 nm. 
     
     
       6. The optics module of  claim 1 , further comprising: a first light source that illuminates the particles in the selection zone, the first light source located on the same optical axis as the first image sensor optical path; and a second light source that illuminates the particles in the detection zone, the second light source located on the same optical axis as the second image sensor optical path. 
     
     
       7. The optics module of  claim 6 , wherein the first light source and second light source emit infrared light. 
     
     
       8. The optics module of  claim 1 , wherein the fluorescence detector assembly comprises a plurality of fluorescence detectors, each of the plurality of fluorescence detectors comprising a silicon photomultiplier (SiPM). 
     
     
       9. The optics module of  claim 1 , further comprising a processor connected to the first image sensor, the second image sensor, the fluorescence detector assembly and the plurality of lasers. 
     
     
       10. The optics module of  claim 1 , wherein the fluorescence detector comprises a photon detector. 
     
     
       11. The optics module of  claim 1 , wherein the optics module is positioned adjacent to a first fluidic channel of a fluidic device. 
     
     
       12. The optics module of  claim 2 , wherein the plurality of wavelengths is selected from within a range spanned by 405 nm and 680 nm. 
     
     
       13. The optics module of  claim 9 , wherein the processor estimates a mean fluorescence for the plurality of detectors. 
     
     
       14. A system for selective microfluidic particle processing, the system comprising: a microfluidic chip having a chip inlet, a detection zone, a selection zone and a chip outlet; a processor; a pressure pulse generator module connected to the processor, the pressure pulse generator module constructed to produce a pressure pulse to the chip inlet and to the chip outlet; an optics module connected to the processor, the optics module comprising: a first image sensor that defines a first image sensor optical path that intersects the selection zone, wherein: the first image sensor optical path comprising an objective with a numeric aperture of less than 0.3; and the first image sensor is constructed to capture images of the particles in the selection zone; a plurality of lasers that define a laser optical path that intersects the detection zone, the plurality of lasers constructed to induce fluorescence excitation in the particles; a fluorescence detector assembly that defines a fluorescence detector optical path that intersects the detection zone, wherein: a portion of the fluorescence detector optical path is along the laser optical path; and the fluorescence detector assembly is constructed to detect the fluorescence excitation in the particles; a second image sensor that defines a second image sensor optical path that intersects the detection zone, wherein a portion of the second image sensor optical path is along the laser optical path; the second image sensor optical path comprises an objective with a numeric aperture of greater than 0.3; and the second image sensor is constructed to capture images of the particles in the detection zone, wherein the processor receives data from the optic module, processes the data and actuates the pressure pulse generator module based on processed data; and wherein the detection zone is separated from the selection zone by a delay line, wherein the magnitude of the delay line is sufficient to allow:
 (1) the processor to process the data received from the optics module; 
 (2) the processor to actuate the pressure pulse generator module; and 
 (3) the pressure pulse generator module to create a pressure pulse at the chip outlet. 
 
     
     
       15. The system of  claim 14 , wherein the fluorescence detector assembly comprises a plurality of fluorescence detectors and a plurality of filters constructed to detect the fluorescence excitation in the particles at a plurality of wavelengths. 
     
     
       16. The system of  claim 15 , wherein the plurality of wavelengths is selected from a group consisting of: 405 nm, 452 nm, 525 nm, 600 nm, and 680 nm. 
     
     
       17. The system of  claim 14 , wherein the plurality of lasers emit laser light at a plurality of wavelengths. 
     
     
       18. The system of  claim 14 , further comprising: a first light source constructed to illuminate the particles in the selection zone, the first light source located on the same optical axis as the first image sensor optical path; and a second light source constructed to illuminate the particles in the detection zone, the second light source located on the same optical axis as the second image sensor optical path.